Biomedical subjects

Chan-Wha Kim

Publications and source records attributed to Chan-Wha Kim.

At least 37 records · Page 2Linked to original sources

PubMed2004

Differential expression of proteins in rat plasma exposed to benzene.

Benzene, a ubiquitous environmental contaminant, is an important solvent in the chemical industry and is also known as a constituent of petroleum. It has been reported that benzene is associated with hematotoxicity including leukemia in humans and cancer in laboratory animals. To study protein expression alterations in rat plasma exposed to benzene, rats were exposed to levels of 1, 10, 100 ppm benzine for 6 h/day and 5 d/week for 2 or 6 weeks. Two-dimensional gel electrophoresis of rat plasma was carried out, and approximately 1000 protein spots were detected on the gels. The 11 spots which showed significantly different expression were selected and identified with matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. Analyzing the targeted 11 spots, there was no correlation between the 2 and 6 weeks benzene-inhaled groups on up-regulated proteins (zinc finger protein, and tristetraprolin) and on down-regulated proteins (cAMP-regulated guanine nucleotide exchange factor II, protein kinase and unknown protein). The overexpressed proteins (inhibitor of kappaB-like protein, GTP-binding protein rab14, T-cell receptor alpha chain, and somatostatin transactivating factor-1) were detected only in groups inhaling benzene for 6 weeks. Among them the expression level of T-cell receptor alpha chain was confirmed by Western blot.

Animals↗

PubMed2004

Insulin microcrystal suspension as a long-acting formulation for pulmonary delivery.

Pulmonary delivery provides the most promising non-parenteral route of insulin administration. Insulin was used as a model protein to demonstrate the feasibility of using protein crystals for the pulmonary delivery of a sustained-release protein drug formulation. Insulin microcrystals with a mean diameter of 3 microm were prepared using a seed zone method. The yield of crystallization was very high (95.8 +/- 0.97%), and the microcrystals were recovered with high efficiency (>98%) by centrifugation. Morphological examination using scanning electron microphotography showed the microcrystals to be of a homogeneous rhombohedral shape, with some rhombus forms, without aggregates. After the administration of 32 U/kg of the microcrystal suspension to STZ-induced diabetic SD rats by intratracheal instillation, the blood glucose levels were reduced and hypoglycemia was prolonged over 13 h, as compared to the insulin solution. The percent minimum reductions of the blood glucose concentration (% MRBG) produced by the microcrystal suspension and insulin solution reached 36.5 and 37.2%, respectively, of the initial level, and the percent total reductions in blood glucose (% TRBG(13 h)) were 34.4 and 25.0%, respectively. In the case of inhalation using a sieve-type ultrasonic nebulizer, the % MRBG produced by the microcrystal suspension and insulin solution were 21.7 and 26.3%, respectively, of the initial level, and the % TRBG(13 h) were 66.7 and 58.4%, respectively. However, the hypoglycemic effects of the microcrystal suspension were prolonged over 7 h, which compares favorably with the insulin solution (P<0.5 by unpaired t-test). These results could be attributed to the sustained-release of insulin from the microcrystals, which were deposited widely throughout the entire lung.

Administration, Inhalation↗

PubMed2004

Analysis of differentially regulated proteins in TM4 cells treated with bisphenol A.

BPA, bisphenol A, a monomer of epoxy resins and polycarbonate plastic, is used in many consumer products including the plastic linings of cans for food and babies' bottles. BPA has been reported to cause reproductive toxicity and affects cells in rats and mice at high doses. In this study, the effect of BPA on protein expression in TM4 cells (a mouse Sertoli cell line) known to play an essential role in Spermatogenesis was investigated by two-dimensional electrophoresis (2-DE). After 16 h exposure to 50, 100, 150, 200, and 250 microM of BPA, the viability of TM4 cells decreased to about 90, 85, 78, 55, and 30% of control respectively. Approximately 800 protein spots in TM4 cells were analyzed by 2-DE with pH 4-7 linear immobilized pH gradient (IPG) Dry Strip, and 11 proteins which showed significantly different expression levels were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Among these, HSP 27 and placental calcium binding protein may be proteins differentially expressed by BPA exposure.

Animals↗

PubMed2004

Microencapsulation of insulin microcrystals.

Insulin microcrystals were encapsulated (microcrystal/PLGA) within poly(lactide-co-glycolide) (PLGA 50:50) by the multiple emulsification solvent evaporation technique and compared with insulin solution microspheres (solution/PLGA) in terms of their morphology, size distribution, drug content, encapsulation efficiency, and stability of insulin during release.

Capsules↗

PubMed2003

Enhancement of mouse pancreatic regeneration and HIT-T15 cell proliferation with rat pancreatic extract.

In this study, the effects of rat pancreatic extract (RPE) on regeneration of impaired mouse pancreas and proliferation of beta-cell line (HIT-T15) were investigated. RPE from the regenerating pancreas (2 days after 60% pancreatectomy) was treated to cure streptozotocin (STZ) induced diabetes in BALB/c mice. RPE-treated BALB/c mice for 21 consecutive days became euglycemic by day 30 and remained normoglycemic during a 150 day follow-up. Saline treated mice remained hyperglycemic sustained uncontrolled hyperglycemia. Islet neogenesis was observed in RPE-treated mice and confirmed by use of immunocytochemistry. Morphometric analysis of pancreas in reverted RPE-treated mice showed a new population of small islets compared with saline controls and an increased islet number. When HIT-T15 cells were treated with RPE, HIT-T15 cell proliferation and insulin secretion increased with increases in the RPE concentration. These results imply that RPE have the regeneration factors and help in the cure of diabetes.

Animals↗

PubMed2003

Microcrystallization of indomethacin using a pH-shift method.

This study developed a microcrystallization process for indomethacin, a nonsteroidal anti-inflammatory drug (NSAID), using a pH-shift method in aqueous solution. The physicochemical properties of the microcrystals produced were similar to those of the standard crystalline powder in X-ray diffraction (XRD), differential scanning calorimetry (DSC), and Fourier transform infrared spectroscopy (FT-IR) analyses, except for a lower XRD peak height and a slightly lower melting temperature (Tm) (1.5 degrees C). Phase contrast microscopy and scanning electron microscopy (SEM) showed that the indomethacin microcrystals were plate-like with a uniform size distribution (mean diameter =10.4+/-0.4 microm). In the initial phase, the dissolution rate of the indomethacin microcrystals was about 2.2 times higher than that of the standard crystalline powder. The biological activity of the indomethacin microcrystals was about 20% higher than that of the standard crystalline powder in their ability to inhibit the proliferation of colon cancer cells (HT-29).

Cell Survival↗

PubMed2003

DNA damage in T- and B-lymphocytes and granulocytes in emission inspection and incineration workers exposed to polycyclic aromatic hydrocarbons.

In this study, we investigated by using comet assay the effects of polycyclic aromatic hydrocarbon (PAH) as a major factor on DNA damage of workers exposed to exhaust fumes. Twenty-four workers from three automobile emission inspection companies, 28 workers from a waste incinerating company, and 43 matched, unexposed healthy subjects were enrolled in the study. The mean values of 1-hydroxypyrene (1-OHP) in automobile emission inspection and waste incineration workers were 0.27+/-0.19 and 0.57+/-0.46 micromol/mol creatinine, respectively, and the mean values of 2-naphthol in automobile emission inspectors and waste incineration workers were 4.80+/-4.01 and 8.30+/-4.79 mol/mol creatinine, respectively. Significant difference in urinary metabolites, 1-hydroxypyrene and 2-naphthol was found between smokers and non-smokers in exposed groups and it may be due to the amounts of smoking cigarettes. In T-lymphocytes, DNA damage in control subjects, emission inspection workers and incineration workers were 1.42+/-0.22, 1.41+/-0.22 and 1.76+/-0.27, respectively. DNA damage of B-lymphocytes in the three groups showed the most significant differences of three cell types. The tail moments of the B-lymphocytes of control subjects, emission inspection and incineration workers were 1.40+/-0.27, 2.44+/-0.32 and 2.36+/-0.37, respectively. In granulocytes, DNA damage was also different, the tail moments being 2.72+/-0.59, 3.32+/-0.38 and 2.85+/-0.49, respectively. Although 1-OHP and 2-naphthol levels were statistically increased in smokers in workers exposed to PAHs, exposed smoking and non-smoking workers did not show any significantly difference in terms of Olive tail moments. Our results suggest that PAH causes single strand DNA breakage in human T- and B-lymphocytes, and granulocytes. A comparison of DNA damage in three groups showed that B-lymphocytes are useful target in the biomonitoring of human exposure.

Adult↗

PubMed2003

Improvement of the two-dimensional gel electrophoresis analysis for the proteome study of Halobacterium salinarum.

Inherent problems exist in the use of two-dimensional gel electrophoresis (2-DE) for sample preparation and separation of proteins from Halobacterium salinarum. In particular, proteins from cells grown in 25% NaCl are difficult to resolve by 2-DE due to the abundance of salt. To remove salts, a 3 kDa molecular weight cut-off column was used. When soluble proteins were separated by 2-DE, most of the proteins were concentrated in the acidic range. For separation of proteins in the pH 3-6 range, ultrazoom immobilized pH gradient strips were used. In addition, sample separation using a IPGphor/Multiphor combined system was a more effective method for the proteome analysis of acidic proteins than using IPGphor for the isoelectric focusing step.

Databases, Protein↗

PubMed2003

Analysis of proteins expressed in rat plasma exposed to dioxin using 2-dimensional gel electrophoresis.

Dioxins are a class of polyhalogenated aromatic hydrocarbons that induces a wide spectrum of toxic responses in animals. In this study, two groups of Sprague-Dawley rats were exposed to 2,3,7,8-tetrachlorobenzo-p-dioxin (TCDD); one group received short-term exposure at a single dose of 1, 10, 20 or 50 microg/kg body weight and the other received long-term exposure to a daily low dose of 0.01, 0.1, 1 or 2.5 microg/kg body weight for one month. Two-dimensional electrophoresis was utilized to resolve the protein profile of rat plasma exposed to TCDD at different doses. One novel and three volume-increased spots were identified and characterized by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and electrospray-ionization on quadropole-TOF2 mass spectrometry. The novel protein was identified as plasma glutathione peroxidase precursor and the volume-increased proteins were cytokeratin 8 polypeptide, Ig lambda-1 chain C region and Ig lambda-2 chain C region. These proteins may be used as biomarkers to diagnose dioxin exposure and may help in understanding the toxic effects of dioxins.

Amino Acid Sequence↗

PubMed2003

Monitoring protein expression by proteomics: human plasma exposed to benzene.

Low levels and long term exposure to benzene is associated with hematotoxicity including aplastic anemia, acute myelogenous leukemia, and lymphoma. Current biomonitoring methods such as urinary phenol, S-phenylmercapturic acid, and trans-trans muconic acid were found to be unreliable as analytical methods to detect benzene exposure. Therefore, to search for a specific protein for biomonitoring benzene exposure, we investigated plasma proteins from workers (n = 50) at a printing company who were exposed to benzene, by two-dimensional gel electrophoresis. The protein profiles are significantly different (p < 0.05) between benzene exposed and unexposed groups, as identified by matrix-assisted laser desorption ionization/time of flight mass spectrometry and confirmed by Western blot analyses. T cell receptor beta chain (TCR beta), FK506-binding protein, and matrix metalloproteinase-13 were expressed only in benzene exposed workers. In addition, interleukin-4 receptor alpha chain and T cell surface glycoprotein CD1b precursor were found to be up-regulated in the plasma of benzene exposed workers. When we treated Jurkat cells with benzene (10 microM-10 mM), TCR beta expression was increased in the membrane more than 6-9-fold compared to untreated cells. In addition, the amount of TCR beta released into the culture media, at benzene concentrations greater than 50 microM, increased up to 10 mM. Therefore, TCR beta levels in plasma could be used as a biomarker and a possible therapeutic target for benzene exposure.

Benzene↗

PubMed2003

Development of an effective sample preparation method for the proteome analysis of body fluids using 2-D gel electrophoresis.

A sample preparation is still the most critical step in two-dimensional electrophoresis (2-DE) and should be optimized for each type of sample. In this study, a protein extraction method from body fluids was developed using a combined centrifugal filter device and a sample treating buffer. When plasma, amniotic fluid, urine, and tear were tested with this method, the recovery of protein reached almost 90% and high-quality separation of 2-DE gel was obtained.

Amniotic Fluid↗

PubMed2003

Characterization of streptozotocin-induced diabetic rats and pharmacodynamics of insulin formulations.

Morphological and functional changes of rat pancreatic islets caused by administration of streptozotocin (STZ) and the bioavailability of insulin formulations administered to STZ-induced diabetic rats with fasting (12 h) or non-fasting were investigated. Islets isolated from normal rats maintained a good three-dimensional structure and the islet yield was 962.5+/-86.5 islet equivalent number (IEQ, islets converted to an average diameter of 150 microm). In the diabetic group (>500 mg/ml blood glucose), the islet yield was only 44.4+/-8.3 IEQ and the islet was severely damaged. The minimum reduction of blood glucose of each formulation, such as insulin solution, microcrystal, and insulin microcrystal capsule, was shown to be 11.3, 11.0, and 16.3 mg/dl, respectively, at 6 h in fasting with diabetic rats. These results indicated that the administration of insulin formulations to the fasting groups increased the severe hypoglycemic effect of insulin action more than in non-fasting diabetic rats. The diabetic rat with fasting has a regulatory disorder in maintaining the blood glucose level. Accordingly, the validity of pharmacological availability as an optimal modeling of insulin formulations is best in non-fasting STZ-induced diabetic rats.

Animals↗

PubMed2002

Ethanol decreases basal insulin secretion from HIT-T15 cells.

Various epidemiological studies suggest that alcohol intake is one of the risk factors leading to type II or non-insulin-dependent diabetes mellitus (NIDDM), but the effect of alcohol on beta-cell function remains unexplored. To study the mechanism of the diabetogenic action of ethanol, we investigated the effect of ethanol on beta-cell functions using a single clonal beta-cell line, HIT-T15 cells. When HIT cells were treated with ethanol, the metabolic activity judged by MTT assay was inhibited in dose- and time dependent manners, but cytotoxicity was not observed. Ethanol also inhibited basal insulin secretion by 30% compared to the untreated control. However, glucose-stimulated insulin secretion was not impaired by ethanol although the basal insulin secretion was inhibited. These results imply that ethanol exert beta-cells to overwork in order to compensate inhibition of the basal secretion. This finding may at least in part explain the diabetogenic action of ethanol.

Cell Line↗

PubMed2002

High-level TNF-alpha secretion and macrophage activity with soluble beta-glucans from Saccharomyces cerevisiae.

We have previously reported that water-soluble beta-glucan completely devoid of mannoprotein and purified from the yeast cell wall effectively stimulated the macrophage function (Biosci. Biotechnol., Biochem., 65, 4, 837-841 (2001)). In this present study, to increase the yield of water-soluble beta-glucan, the wild type of Sacharomyces cerevisiae, JH, was treated with a combination of UV irradiation and laminarinase (endo-beta-(1,3)-glucanase) to yield the laminarinase-resistant mutants, JUL1 and JUL3. Water-soluble beta-glucans that were free of mannoprotein from JH, JUL1 and JUL3 were purified and their effects on TNF-alpha secretion and phagocytosis by macrophages were evaluated. Crude beta-glucan was first solubilized from the yeast cell wall by alkaline extraction and then subjected to an acid treatment. The residual mannoprotein was completely removed by DEAE and ConA chromatography. The yield of water-soluble beta-glucan in both mutants (JUL1, 5.11%; JUL3, 5.76%) was about 5-fold higher than that of the wild type (1.16%). The water-soluble beta-glucan from JH induced TNF-alpha secretion slightly more than that from JUL1 or JUL3: TNF-alpha secretion by JH at 50, 200, 500 microg/ml of beta-glucan was 11-17% more than that by JUL1 or JUL3 for the same treatment. Beta-glucan from the wild type stimulated phagocytosis slightly more than that from the mutants. These mutants could therefore effectively produce purified water-soluble beta-glucan with immune activity.

Animals↗

PubMedDate not supplied

Plasma protein level changes in waste incineration workers exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin.

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) is a chemical compound which is known to induce severe reproductive and developmental problems, immune system damage, and interference with regulatory hormones. To characterize changes in the expression of plasma proteins caused by exposure to TCDD, we analyzed plasma samples from workers at municipal incinerators using two-dimensional gel electrophoresis (2-DE). Proteins exhibiting differences in expression were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) and electrospray ionization quadrupole (ESI-Q) TOF mass spectrometry. One newly expressed protein was identified as the adrenomedulin binding protein (AMBP). Seven overexpressed proteins were identified in this study, and the most overexpressed protein was identified as alpha-fetoprotein (AFP). In addition, we cultured HepG2 cells in the presence of TCDD, to determine the effects of TCDD on the AFP and albumin expression in mRNA and protein levels, via RT-PCR and Western blotting, respectively. TCDD treatment resulted in an increase in the mRNA and protein expression levels of AFP, but reduced albumin expression. According to our results, exposure to TCDD may induce liver disease or cancer, and the proteins identified in this study could help reveal the mechanisms underlying TCDD toxicity.

Albumins↗

PubMedDate not supplied

Comparative analysis of the tear protein expression in blepharitis patients using two-dimensional electrophoresis.

Change in the expression of body fluid proteins is caused by many diseases or environmental disturbances. The changes in tear proteins are also associated with various pathological eye conditions. Especially, chronic blepharitis is one of the most common conditions seen in the ophthalmologist's office. However, there are no specific clinical diagnostic tests for blepharitis, and it is difficult to treat effectively. Therefore, the aim of this study was to screen prognostic or diagnostic marker tear proteins for blepharitis and investigate pathogenesis of this disease using proteomics techniques. The tear proteins expressed in patients suffering from blepharitis (patient, n=19) and healthy volunteers (control, n=27) were analyzed using the two-dimensional electrophoresis (2-DE) technique. The differentially expressed proteins in patients were identified with ESI-Q-TOF (electrospray-quadrupole-time-of-flight) mass spectrometry and confirmed with western blotting. Nine proteins in patient were down regulated about 50% compared to those of the control: serum albumin precursor, alpha-1 antitrypsin, lacritin precursor, lysozyme, Ig-kappa chain VIII, prolactin inducible protein (PIP/GCDFP-15), cystatin-SA III, pyruvate kinase, and an unnamed protein. The use of the two-dimensional eletrophoretic technique could give more insight into the disease-related protein expression changes in tear fluids. Our findings reveal that the composition of tear proteins in blepharitis patients is different from that of healthy subjects and may provide further insights into the pathogenesis of blepharitis.

Aged↗

PubMedDate not supplied

Proteomic characterization of rat liver exposed to 2,3,7,8-tetrachlorobenzo-p-dioxin.

Dioxins are a class of polyhalogenated aromatic hydrocarbons that induce a wide spectrum of toxic responses in experimental animals. In this study, 2,3,7,8-tetrachlorobenzo-p-dioxin (TCDD) was exposed to two SD rat groups; one group for short-term exposure at a single dose of 1, 10, 20 and 50 mug/kg body weight (group 1) and the other for long-term exposure at daily and-low dose of 0.01, 0.1, 1 and 2.5 microg/kg body weight (group 2) for a month. Two-dimensional electrophoresis (2-DE) was utilized to resolve the protein profile of rat liver exposed to TCDD at different doses. In the analysis of 2-DE of the group 1, two new-expressed spots and seven volume-increased spots were detected and identified by ESI-Q-TOF MS/MS; especially, proteasome subunit beta type 3 was increased in all doses. In addition, in the group 2, six volume-increased spots were screened; particularly, histidine triad nucleotide binding protein was increased in both 0.1 microg/kg dose and 1 microg/kg dose. The identified proteins were confirmed using Western blot. Among the identified proteins, apolipoprotein A-IV may protect lipid peroxidation and atherosclerosis induced by TCDD exposure and the expression level of phosphoglycerate mutase increases due to hyperthyroidism induced by TCDD exposure.

Amino Acid Sequence↗

PubMedDate not supplied

Changes in the cell size of Brevundimonas diminuta using different growth agitation rates.

Brevundimonas diminuta (ATCC 19146) is a standard organism for validation of sterilizing-grade membrane filters. Cell size is critical for the determination of retention characteristics of 0.2 micron rated membrane filters. In this study, cell size changes of B. diminuta cultured under different physiologic states and variable agitations at 50, 100 and 200 rpm were measured by a particle size analyzer and scanning electron microscope (SEM). The smallest cells were obtained at initial stationary phase in saline lactose broth (SLB) as a shaking culture at 50 rpm. Cells grown under agitation at 50, 100 and 200 rpm showed an increase of specific growth rate (mu), about 2.9, 3.6 and 3.6 fold, respectively, compared to the non-agitated cells in SLB media. These results suggested that the cell size decreased proportionally with increase of the specific growth rate (mu) in SLB. These size changes were associated with penetration through a 0.2 micron rated cellulose acetate filter. A scale-down filtration system was developed and performed bacterial challenge test and bubble point test with cells cultured in SLB. Cells grown under agitation conditions in SLB were not retained by 0.2 micron rated membrane filter.

Bacteriological Techniques↗